MicrobiologicalInvestigationofOrganicManures. 1. DecompositionofRape-cake. By .AraoItanoandSatiyoArakawa. [00ωber 10 , 193 勾 Th i 日 paperpre 自ent 自 ther 掛 u1tsobtainedon 也edecompOBitionofrap e-c ake by two回 parateinv 舗もig a. tion 日, n a.m e1yParlA. Decomposition ofr a. pe-cake in 日 oi 1s and ParlB. Decomposition of rape -c akeby various knownstock culture 目. Th e r a. pe ・ cakei 自 oneoftheorga 凶cmanure 白 whichareused戸 rticu1ar1yin Japan , incultivatingcerlaincropssuch幽 tobacco , omnges , ahrubsandother orn a.m ental fl. owers , and it is con 日 ideredtohavecertainmeritRtoproducebetter cropsinreg a.r d to fl. avor , co1ourandtastebesid 伺 havingtheu 自 u a. 1nitrogen supp1y. Os UOI岨 d YOSIE I) underlookagenem1inv 倒 tigationon也 ed 舵 ompositionof org 岨 icnitrogenousmaDuresin 自oilunderdi 宜 erenttemperature , moistureand 的i1 texture , and found thatthempe-cakeis出emos 七rωistantsubs も aDcetothe d 舵 omposi “ on. 宜O ElI MURA andhi 自 cc トworkers 2 ) investig a. tedthempe ・ caketo制 certain出e causal constituentswhich may bere 自 ponsib1ein making the mp e- cake 剖 thebet も ermanurefortobaccoandothercertain vegetablesも hanotherorga 凶O manure such a 自由oybe 岨 cake. Th ey found 也 attherawrape ・ cake ∞ ntainsmore choline than that 白 thesoybean cake , and inthedecomposedrape -c ake , a 1a rger quantity of 1actic acid , 1eucin 阻 dmust 町 doil IB produced , a. nd801 岡山eyfound 也atthe decompo 白 ,itionproce 目白 i 日very 日 10wmaitaining 也eacid reactionthrough- out も heproce 自由 andproducemuchorganic 白色1t s 踊 theintermediateprαluc 旬 w hi1 e the quantity of ammonia decre 幽飢 A 自 tothefactorswhichiD fl. uencethedecomposi “ on of organicmaDures , m 岨 y investig a. tion 自 h a. ve beenuDdertakenand known that the proce 闘 i 自 infI. uenced verym 町 'ked1y. on 出 e0 也 erhand , very litt1ei 自 known a. bout 白 emicroorg 岨 ISIDswhichmaybringaboutthedecomp 佃 ition.Consequen t1 y it w 朗 inve 自tigatedfirst 朗 tothe kinds of m Ul r ∞ rganisms whichmaybring about the a.m monmcation whi1e the r a. pe -ca ke underg 伺自也edecomposit おninthe 回 i1 , and later by the known cultures.
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Microbiological Investigation of Organic Manures.
1. Decomposition of Rape-cake.
By
.Arao Itano and Satiyo Arakawa.
[00ωber 10, 193勾
Thi日paperpre自ent自 ther掛 u1tsobtained on也edecompOBition of rape-cake
by two回 parateinv舗もiga.tion日, na.me1y Parl A. Decomposition of ra.pe-cake
in 日oi1sand Parl B. Decomposition of rape-cake by various known stock
culture目.
The ra.pe・cakei自 oneof the orga凶cmanure白 whichare used戸rticu1ar1yin
Japan, in cultivating cerlain crops such幽 tobacco,omnges, ahrubs and other orna.mental fl.owers, and it is con日ideredto have certain meritR to produce better
crops in rega.rd to fl.avor, co1our and taste besid伺 havingthe u自ua.1nitrogen
supp1y.
OsUOI岨 dYOSIEI) underlook a genem1 inv倒 tigationon也ed舵 ompositionof
org岨 icnitrogenous maDures in自oilunder di宜erenttemperature, moisture and 的i1texture, and found that the mpe-cake is出emos七rωistantsubsもaDceto the
d舵 omposi“on.宜OElIMURAand hi自 ccトworkers2)investiga.ted the mpe・caketo制 certain出e
causal constituents which may be re自ponsib1ein making the mpe-cake剖
the betもermanure for tobacco and other certain vegetablesもhanother orga凶O
manure such a自由oybe岨 cake. They found也atthe raw rape・cake∞ntainsmore
choline than that白 thesoybean cake, and in the decomposed rape-cake, a 1arger
out もheproce自由 and produce much organic 白色1ts踊 the intermediate prαluc旬
whi1e the quantity of ammonia decre幽飢
A自 tothe factors which iDfl.uence the decomposi“on of organic maDures, m岨 yinvestiga.tion自 ha.vebeen uDdertaken and known that the proce闘 i自
infI.uenced very m町 'ked1y. on出e0也erhand, very litt1e i自 knowna.bout
白emicroorg岨 ISIDswhich may bring about the decomp佃 ition. Consequent1y
it w朗 inve自tigatedfirst朗 tothe kinds of mUlr∞rganisms which may bring about
the a.mmonmcation whi1e the ra.pe-cake underg伺自也edecompositおnin the回 i1,and later by the known cultures.
428 A. ITANo and S. ABAlUWA:
Part A. Decompωition of Rape-伺kein Soils.
In this invωtigation, the microbiological decomposition of出erape-cake w朗
undertaken in出e同oil自ofwhich conditions imitaもedもhoseof出epaddy-field and
dry-farm following the procedure which will be de日cribedbelow ;
The soils from both the paddy-fteld and dry-farm of our experiment&l
plo句 wereu自edafωr也eyare dried and shuted through 0.5 mm. sieve.
Two grams of rape・品kewhich con也ins5.3% niもrogen,were added to 200 grams of th白 田ilin a widEトmouthedboもtle,and mixed thoroughly, and in case of the paddy-field soil, 72% water and 42%句出自 dry-farmsoil were added
r伺 pecLively加 imit叫ethe natura1 moisture content in e郎 hcase. The bottles
thus prepa.red, were sもopperedwiもh出erubber corks which were provided
with glass tubes ha.ving the cotton p1ugs, and pla.ced in an incubator 叫 2800.
On tbe fourth day, no apparent change WIl.S ob3erved buも色hera.pe・cakeculture
gave a pecu1iar odor which resemb1e自由叫 of出eactinomyces. After旬nd島ys
The moi目tureconもentw制 determinedby the standard me出od;the ammonia
W幽 determinedby HA.HPE晶、 method8)剖 follow自;15g.日0江 with2∞cc. of 20%
KOl w幽 placedin shak凶gbottle and shaken for 30 minutes and filtered, and 2似トー250cc.of出efi1trate wi出 1g. MgO a.nd a sma.ll pi白巴eof para鑑nw朗 putin
a. Kjeldahl fiask a.nd distilled ;也edis凶1a旬 W剖 collectedin加 N/l∞H2S0,with meもhylred indicator; after driving ou七OOlby boiling, titrated with N/l∞ Na.OH. The ni色mも争nitrogenw倒 determined by placing 10 g. 自oilin 5∞cc.
Erlenmeyer fta.sk with 1∞c巴・ wa.tera.nd sbaken for 20 minutes, and副総red
wi出品目malla.mounもofCaO a.nd the aolorim白色ricmethod was a.pplied by using
phenol-di・sulpho凶0加 id闘 usual.
The number of bacteria w剖 determinedby W AKSMA.N'S me出odのbyta.king 1-1ω,似ぬ or1-10刈凧αX>dilution, and using出efollowing medium :
Glu巴O自e
KzHP04 MgS04
1.0g.
0.5g.
O.2g.
FeS04 仕a.ce.
Egg a.lbumin (powdered) 0.25 g.
H20
Agar
1,∞Occ.
1.5.5g.
The colonies were coun旬dafter ten days incubation at 2SOC.
Mio同 biologioal1nv曲tJgationof Orgnnio M岨 U回 s. 1. 429
DelerminaJion o.f Inorg,側 ri'cN;lrog側 Z
The quantity of inorganic nitrogen produced is given in Table 1 IIond
The resul旬由ownin Ta.ble II a.nd Figure II resemble those which were given in Ta.ble 1 a.nd Figure 1. Tha.t is the number of ba.c旬ria.in the pa.ddy-field
soil increa.sed ra.pidly on the fourth da.y up加 出et自ntha.nd stea.dily incre幽 ed
thereon. While the dry-fぽ m soil showed the sa.meωndency but more in初 旬l
num ber a.nd the ma.ximum ca.me on th自twentyfifth da.y a.nd decl'ea.sed somewh叫la.ter.
JforthologJ'臼'.]Groutl飢Ig0/ Bacleria ..
The ba.cteria. were grouped morphologica.lly into Euba.c飴ria.lestype a.nd
N.>le3: G. = gll10l3e j QC}. = qlllloDtity of glnolse j 8pN. = 801uble proteln nitrogen.
The a.bove r剖 ult8indica.teもha.tthe addition of g1uc08e ma.de出自 rea.ctionof
medium a.cid朗出eculture grew a.nd 80me were cha.nged句 le聞もha.nPH 3.0 which 泊 letha.lrea.ction for the org晶nis乱 Theacidifica.tion i8 evidantly .ca.u8ed by
the production of orga.nic acid from glucose but it was noもinp品目11elwith the
a.mounもofglucose consumed. The ba.cteria. a.nd 80me a.ctinomyces con8umed
very sma.ll amount of gluc伺 ewhile出efungi a.nd certa.in a.ctinomyce8 co田 umed
markedly. The a.ddition of glucose ill1luences the genera.l a.pp叫 ra.nceof ra.pe-ca.ke de巴omposition制色o他。 ωlora.tionof o;tedium and the vigor of mycelium
grow出 offungi a.nd a.ctinomy日倒・ After 1) da.Yi!, no NHl-N w嗣 detecもedin 11011
the cultures except 110 slight production in the fungi, a.nd a.lso less tha.n ha.lf of
decomposition of出era.pe・叫ke,阻 dwa.s found tha.t the actinomyces a.s a. who1e
wa.s the strongest阻lOngthese microorg阻 ismo.
2) Among th白b附 eria.,出espor白 formerswere stronger tha.n the non句 ore
formers. B. my∞ides, B. cen旭, B. tumωcens a.nd B. mega.出自riumwere the
strong ones a.mong the former a.nd B. prodigi佃 usw踊 rela.tive1ystrong a.mong the 1a.悦er.
3) Among the fungi, Asper. cellu10s剖, Asper. oryza.e,αadsporium herba.rum
and Tl'ichoderma. koni企giiwere ra.ther strong. A1so Rhizopus nigrica.n自(?)which
is common1y found on th自 ra.p刊誌ew鍋 fa.ir1y町 ongdecomposer. Different
stra.in of fungi w踊 differentin their power of decomposition.
4) In genel'sl, the sa.nd culture ga.ve better d郎 ompositionもha.nthe 1iquid
cuILure.
5) The ~mecha.nism of rape-ca.ke decompo目itionseems to be tha.t也esolub1e
non-proωin Bubst岨 ceis produced first a.nd gradua.lly cha.ngedもoa.mmonia.. on the fifth dsy, the fo即時rw嗣 foundmo日ta.bundsntly a.nd it w剖a.fterfif凶ento twenty da.ys when the 1a.tt自rwa.s produced in qusntity. The ma.ximum produc-
tion of solub1e tota.1 nitrogen w嗣 rescheda.t旬nor fifteen da.ys. As a. ru1e tho随
stra.in目whichproduωd a.bunda.nt solub1e protein nitrogen showed a. small ra.tio
in soluble加ω1niもrogen: NHa-N, a.nd it w朗 veryma.rked inωse of出e
恥 tinomyc倒.
6) A slight回 idrea.ction a.t the ihitia.1自ta.geor PHく7w制 bettersuited for
the ra.pe・ca.kedecomposition.
7) An a.ddition of 2% glucose句 themedium, ID8l'k伺 thereaction of
medium very a.cid and the production of solub1e niもrogenwa.s reducedもoa.1most
one ha.lf of也a.twithout gluco随・ Even a.fter fifteen da戸, it rema.ined the sa.me.
No pa.ra.llel rela.tion between the a.mount of gluω回 consumeda.nd the即 id
production w制 observed.
The a.u出orswi自hto acknow1edge Mr. T. MAT師 MOTO'Sa.ssista.nce rendered
in ma.king ma.ny determina.tions.
446 A. 1T..uro Ilnd B. AlU.XA.wA.:
B eferences :
1) B. OsUGI and B. YOSI1l, J. Agr. Ohem. 800., .T"l皿n,4: 6-9, 1929. 2) K. Y目立MUBA.Qud T. MUBA.TA, Bull. K..gosim島 α)1.Agr. &; Forestry, No. 7, 49, 1929. 3) IU且l'EB, H. J., Boil Boienoe, 18: 409, 1927. 4) B. A. W A.XlIMAN,制1Soienoe, 14: 283, 1922. 5) ibid. 8: 71, 1919; J. &ot., 5: 1, 1920.